University Of Texas At Austin

NIH Research Projects · FY 2026 · 2001-09

PROJECT SUMMARY This project is a continuation and further development of a previous INIA project, which was based on studies showing ethanol-induced changes in neuroimmune gene expression in animal models and humans. Those data suggested that ethanol dysregulates Toll-like receptor (TLR) signaling through the myeloid differentiation primary response gene 88 (MyD88) and thereby promotes excessive ethanol intake. Our most recent work showed that genetic and pharmacological manipulation of another branch of TLR signaling via the TIR-domain- containing adapter-inducing interferon-β (TRIF) protein also regulates ethanol intake. In exploring downstream mechanisms by which these signaling disruptions act, we found that chronic alcohol consumption increased several TRIF-dependent signaling components, including type 1 interferons (IFN1s). These findings lead us to the central hypothesis of this renewal application, which is that chronic alcohol exposure activates pathways leading to IFN1 production and expression of interferon-stimulated genes (ISGs), which increase alcohol consumption. To test this hypothesis, we will study alcohol intake and alcohol-related behaviors in mice deficient in critical components of pathways leading to IFN1 production and signaling, and in mice administered compounds that block IFN1 signaling. The proposal has three Specific Aims. Specific Aim 1 will study changes in alcohol consumption in mice undergoing every other day two-bottle choice (EOD-2BC) drinking for at least 4 weeks. To reduce IFN1 production, we will examine these behaviors in mice with genetic deletion of the transcription factors Irf3 or Irf7. To examine the role of IFN1 signaling, we will use Ifnar1 knockout mice which carry a null mutation in the IFN1 receptor. We will also study wild-type mice treated with inhibitors of the kinase TYK2, which mediates IFN1 receptor signaling. Specific Aim 2 will determine if EOD-2BC drinking induces IFN1 responses in specific brain regions by detecting Fos expression in Mx1GFP mice in which the interferon-stimulated response element of the Mx1 gene drives GFP expression. Data from these whole brain imaging studies will be shared with the Harsan-Keiffer project to be incorporated into their multimodal connectome analyses. To determine if IFN1 regulates alcohol responses in certain regions, we will knockout Ifnar1 using local microinjection of Cre recombinase in floxed Ifnar mice and use Fos-Cre-ER (TRAP2) mice to knockout or knockdown Ifnar1 in activated neurons and glia. Specific Aim 3 will identify ISGs induced by EOD-2BC alcohol drinking in brain regions identified in Aim 2. The Mayfield project will help us analyze transcriptomic data to identify ISGs. To investigate causality, groups of ISGs will be knocked down using multiplex CRISPR interference in collaboration with the Farris-Homanics project. We anticipate that some other INIA projects will identify additional proteins that require behavioral testing with pharmacological agents or genetically modified mice, which we will carry out as part of this consortium. !

NIH Research Projects · FY 2025 · 2001-09

Abstract With the generation of more than 100 sequenced vertebrate genomes, the current key question is how to determine the role(s) of uncharacterized gene products in specific biological and pathological processes. For example, genes associated with human disease are being discovered at a rapid rate, thanks in part to the development of next generation sequencing technologies. However, the biological functions underlying this linkage are often unclear, in part because the majority of published work investigates genes encoded by ~10% of the genome. The overall purpose of this long-standing and collaborative research program is to develop innovative ways to address this key gap of functional annotation in genome science. For example, mitochondria have integral functions in metabolism, organ homeostasis, apoptosis and aging. They also play important but still largely perplexing roles in human pathophysiology, as demonstrated by the enormous biological variation and diverse disorders in patients with mitochondrial disease that can compromise nearly every organ system. Over 1100 known nuclear proteins reside in vertebrate mitochondria, with the majority of unknown biological function or no known role in pathogenesis. Deploying loss of function approaches in model systems has been essential to the annotation of the genome to date from the discovery of novel processes to the biological mechanisms underlying disease. Among vertebrates, Danio rerio (zebrafish) has emerged as an outstanding and pioneering vertebrate amenable to both forward and reverse genetic approaches. This funding period will deploy at scale recently developed genome engineering technologies designed to address these gaps in the field. We will use Predominant Microhomology-mediated end-joining Allele (PreMA) generation for rapid functional screening using reproducible allele generation in F0 animals. For those loci with accessible phenotypes, second phase analyses will follow via GeneWeld large-insert targeted knock-in technology in conjunction with gene-breaking protein traps for detailed functional annotation of the genome using targeted protein trapping in F1+ stable lines. Mitochondria are the products of two genomes – the nucleus and from mtDNA. The conservation between zebrafish and human in the mitochondrial genomes includes nearly identical size and perfect synteny of all 37 protein-coding and RNA genes. We will also deploy our recently developed zebrafish mitochondrial base editor to make the first majority heteroplasmy animal models with targeted mutations in mtDNA-encoded proteins for in vivo functional annotation. We will use this pioneering model organism for mitochondrial genomic annotation focusing initially on the mitochondrially encoded proteome and at a targeted panel of vertebrate-specific, nuclearly encoded conserved mitochondrial proteins of unknown function. We will annotate the functional role of these genes using a rich array of biological, phenotypic and biochemical tests in this highly collaborative research program. This work will yield 1) Novel tools for rapid PreMA deployment and new methods for targeted integration of Gene-breaking Protein Trap alleles will be generated and disseminated 2) New biological and genomic annotation of vertebrate mitochondrial innovations in development and regeneration including mechanisms underlying genetic compensation 3) Longitudinal knowledge of mtDNA SNP heteroplasmy maintenance and 4) Complete a unique functional annotation of the mtDNA-encoded proteome. This combination of novel technologies, annotation and genome science will establish important new functional information on the role of mitochondria in biology that, when compromised, underlies disease.

NIH Research Projects · FY 2025 · 2000-08

Project Summary/Abstract We and others have provided evidence that changes in brain function are caused by neuroadaptations throughout the addiction cycle. Large-scale DNA and RNA sequencing, bioinformatics, and computational approaches have greatly advanced addiction neurobiology. Our newly proposed research will use single cell RNA sequencing to define the transcriptome in human alcoholic and CIE-exposed mouse brain in unprecedented detail. In addition to the similar alcohol-responsive expression changes found in alcoholics and CIE-treated mice, CIE vapor produces escalations in voluntary drinking and is a well-established animal model of alcohol dependence. CIE as well as chronic voluntary consumption models also induce neurobiological and behavioral adaptations in mice that mimic those found in human alcoholics. Unlike whole tissue sequencing, the single cell approach is much more sensitive and prevents dilution of cell-specific expression changes. This proposal will provide the first single cell resolution of convergent genes in human and mouse brain that are associated with alcohol dependence and escalation in drinking. High-priority genes will be functionally validated in mice, and new computational approaches based on integrated single cell-transcriptomic data will then be used to predict and test drugs for efficacy to reduce drinking in mice. Our overarching hypothesis is that conserved cell-type-induced transcriptome changes will reveal specific neurobiological mechanisms and improved drug targets for excessive alcohol drinking. Integrated data from mouse and human brain enhance our ability to predict pathways and drugs with translational relevance in humans.

NIH Research Projects · FY 2026 · 1990-08

This is a year -30 renewal request that addresses a major challenge in medicinal chemistry, namely reducing the systemic toxicity of chemotherapeutics. In prior work, we showed that conjugation of a Pt(IV) center to a water solubilized texaphyrin core called motexfin gadolinium (MGd) allowed for near-complete tumor regrowth suppression in a patient-derived xenograft ovarian cancer mouse model. We now propose to build on this success by creating metallotexaphyrin conjugates bearing poly (ADP-ribose) polymerase 1 (PARP) inhibitors and Au(I) payloads. Advantage will be taken of the fact that the properties of metallotexaphyrins vary with the metal. This, we suggest, will allow us to create tumor-targeting systems with functional features that include an ability to act as so-called PARASHIFT and 19-F nuclear magnetic resonance (NMR) probes, as well as promoting photo-induced thermal and pyroptosis effects. The net result should be systems that not only allow for imaging, but also give rise to therapeutic benefits that are enhanced relative to the active payload alone. To test this hypothesis, we will pursue the following specific aims: 1. Explore paramagnetic water solubilized lanthanide texaphyrins as a new class of potential PARASHIFT probes. Prepare new paramagnetic water soluble texaphyrin complexes and confirm that these complexes produce large downfield or upfield paramagnetic shifts of key 1-H signals (e.g., methyl resonances). Extend these studies to include fluorinated derivatives to allow concurrent 1H- and 19F-NMR based signaling. 2. Prepare and study In vitro texaphyrin conjugates based on PARP inhibitors and Au(I) centers. Synthesize texaphyrin conjugates based on PARP inhibitors and Au(I) centers. Study in platinum sensitive/resistant ovarian, BRCA1 normal/mutated triple negative breast and lung cancer cell lines. Test whether improvements are seen with easier-to-cleave linkers. 3. Test whether cell killing synergies with the conjugated payload are seen under conditions of red- light photo-irradiation in vitro. Test whether damage associated molecular patterns characteristic of pyroptosis or the increases in temperature expected for photothermal effects are seen for key texaphyrin conjugates upon photo-irradiation. 4. Carry the most promising systems on into in vivo studies using murine models. Treat tumor bearing mice with most promising conjugates for desired application i.e., MR imaging, photothermal and pyroptosis. Test acute toxicity, tumor growth inhibition and biodistribution.

NIH Research Projects · FY 2025 · 1987-09

This proposal is for the competing continuation of a T32 training program on the Neurochemical & Behavioral Correlates of EtOH Effects for 5 predoctoral and 3 postdoctoral fellows at The University of Texas at Austin. This multidisciplinary program has a long history of training predoctoral and postdoctoral fellows in innovative research questions related to alcohol misuse to help the national effort of producing the next generation of independent scientists focused on alcohol-related research. Intertwined with the internationally renowned Waggoner Center for Alcohol & Addiction Research, our training environment and opportunities are exceptional, provided by a highly collaborative group of 14 faculty members from seven research units (Behavioral Neuroscience, Clinical Psychology, Molecular Biosciences, Neurology, Neuroscience, Pharmacology-Toxicology, and Psychiatry) and four graduate programs (Cellular and Molecular Biology, Neuroscience, Pharmacology-Toxicology, and Psychology). The primary objective to provide rigorous, state of the art research training includes research in cellular and animal models as well as human subjects and in a breadth of innovative approaches including molecular biology and genetics, bioinformatics, cellular imaging, electrophysiology, neuroanatomy, neurochemistry, human imaging and behavior. Trainees are immersed in the alcohol research field facilitated by the dynamic, intellectual environment provided by the Waggoner Center and the Institute for Neuroscience. Additional objectives include training in effective scientific communication as well as professional and career development. Programmatic elements enhance the experience, including required courses and seminars on Rigor and Reproducibility, Responsible Conduct of Research, Grant Writing, Scientific Communication, and Alcohol Journal Club. Multiple presentation opportunities for trainees are available in a number of seminar series, local symposia, and the T32’s new works-in-progress sessions. Predoctoral students will be required to complete a series of core course requirements, achieve research milestones under direct guidance of training faculty, and satisfy departmental requirements culminating in a Ph.D. degree. Postdoctoral training will be for three years and consist of focused alcohol research guided by a faculty mentor and coupled with professional development activities supported by the training program. Exceptional facilities, resources and commitment to training from all levels provide an outstanding research training environment to prepare our trainees for success in research-intensive careers focused on the causes and consequences of alcohol misuse.

NIH Research Projects · FY 2026 · 1977-07

Summary/Abstract Modified Abstract The Population Research Center (PRC) of the University of Texas at Austin (UT) requests a 9th renewal of its T32 NICHD-funded Training Program in Population Studies. The COVID-19 pandemic, the Dobbs decision reversing Roe, spikes in teen suicide and other threats to youth well-being, and the global increase of displaced persons are just four of the many pressing social issues that affect the health of the U.S. population and call for being addressed through public health and healthcare system solutions. Our training program benefits from an outstanding interdisciplinary faculty with scientific expertise in four key areas of Reproductive Health, Population Health, Family Demography, and Work, Education and Inequality, along with new depth in Biosocial Methods. The PRC provides the infrastructure, training opportunities, and research environment necessary to train the next generation of population scientists and is supported by a strong extramural funding trajectory. PRC predoctoral and postdoctoral fellows are supported by state-of-the-art administrative, computing, scientific, and program development resources; expert faculty collaborators and mentors; topical working groups and a seminar series that provide an intellectual idea exchange and access to well-known scholars from across the country and world. We request training funds for 5 predoctoral fellows and 2 postdoctoral fellows per year for the 2024- 2029 grant period to maintain the size of the program. The center provides an outstanding training environment with notable accomplishments including predoctoral and postdoctoral fellows’ high research productivity in terms of articles and conference presentations, as well as their success in securing external funding for their own work and in securing leading academic, postdoctoral, and research positions around the country. Reflecting the growing interdisciplinarity of population research, our program trains students from Sociology, Human Development and Family Sciences, and Economics, and we expect to continue to draw students from other disciplines, including Psychology and Public Affairs. Through this program, our goal is to continue to support, develop, and produce independent, intellectually engaged scholars from across all demographic groups who produce work of the highest quality and ethical standards. In sum, we request support to continue our success in serving the mission of NICHD’s Population Dynamics Branch to support highly trained, ethical, productive researchers who will inform public health and healthcare delivery solutions to the most pressing national and international public health problems.

Other NSERC · FY 2024

Animal communication, Sexual selection, Behavioural ecology, Frogs, insects, Social network analysis